HO-1 as a diagnostic and prognostic test for dementing diseases

ABSTRACT

The invention relates to a commercial package for determining the concentration of heme oxygenase-1 (HO-1) and/or a nucleotide sequence encoding HO-1 in tissue obtained from a patient, wherein the commercial package is used to predict the onset of, diagnose, or prognosticate a dementing disease. The tissue is suitably plasma, lymphocytes, cerebrospinal fluid or fibroblasts. The commercial package is useful where the dementing disease is any of Alzheimer&#39;s Disease, Age-Associated Cognitive Decline, Progressive Supranuclear Palsy, Vascular (i.e. multi-infarct) Dementia, Lewy Body Dementia, Huntington&#39;s Disease, Down&#39;s syndrome, normal pressure hydrocephalus, corticobasal ganglionic degeneration, multisystem atrophy, head trauma, Creutzfeld-Jacob disease, viral encephalitis and hypothyroidism.

[0001] This application is a division of U.S. application Ser. No.09/323,248, filed Jun. 1, 1999, now allowed, which claims the benefit ofU.S. Provisional Application No. 60/098,141, filed Aug. 27, 1998.

FIELD OF THE INVENTION

[0002] This invention relates to a method for predicting, diagnosing,and prognosticating dementing diseases such as Alzheimer's Disease (AD)and Age-Associated Cognitive Decline (AACD).

BACKGROUND OF THE INVENTION

[0003] Alzheimer's Disease is a neurodegenerative disease which causesdementia. The period from first detection of AD to termination can rangefrom a few years to 15 years, during which time the patientprogressively suffers loss of both mental function and control of bodilyfunctions. There is significant variability in the progress of thedisease. While the majority of patients have a gradual, inexorableprogression (losing on average 3 to 4 points on the 30 point Folsteinmini-mental state score annually), approximately 30% of AD cases have aprolonged stable initial plateau phase lasting several years (Haxby etal., 1992). A subgroup of patients has a fulminant, rapidly progressivedownhill course over several years (Mann et al., 1992). Other patients(about 10% of cohorts) remain slowly progressive, showing only gradualdecline from year to year (Grossi et al., 1988). The pathological,chemical, and molecular bases of this heterogeneity remain undetermined.Recognition of the variability of AD progression represents an importantclinical insight, and may explain the diagnostic difficulties presentedby “atypical” cases.

[0004] Attempts at predicting the onset of AD or monitoring itsprogression have met with limited success. While in certain cases, thereis a familial manifestation of the disease, it appears that the majorityof AD cases are non-familial, and no simple genetic marker for thedisease has yet been determined. Much research has focused on theprotein beta-amyloid, deposits of which are found in the brains of ADvictims.

[0005] Methods of predicting, diagnosing in its very early stage, andprognosticating AD and other dementing diseases are needed.

SUMMARY OF THE INVENTION

[0006] It is a goal of the present invention to provide a method forpredicting, diagnosing, and prognosticating AD and other dementingdiseases.

[0007] In one embodiment, the invention provides a method for assessingdementing diseases in a patient which comprises determining theconcentration of heme oxygenase-1 (HO-1) and/or a nucleotide sequenceencoding HO-1, in tissues obtained from a patient, and comparing saidconcentration with the corresponding concentration of HO-1 and/or anHO-1 encoding nucleotide sequence in corresponding tissues obtained fromat least one control person, wherein such method is used to predict theonset of, diagnose, or prognosticate dementing diseases.

[0008] In another embodiment, the invention provides a commercialpackage comprising means for determining the concentration of hemeoxygenase-1 (HO-1) and/or a nucleotide sequence encoding HO-1, intissues obtained from a patient, and instructions for comparing saidconcentration with an established standard of the correspondingconcentration of HO-1 and/or an HO-1 encoding nucleotide sequence incorresponding tissues obtained from at least one normal age-matchedcontrol person or from the patient at an earlier time.

[0009] The tissues to which the method can be applied include plasma,cerebrospinal fluid, lymphocytes and fibroblasts.

[0010] Advantageously, embodiments of this invention provide an easilyadministered blood or cerebrospinal fluid test which is used to predict,diagnose, or prognosticate AD and other dementing diseases.

BRIEF DESCRIPTION OF THE DRAWINGS

[0011]FIG. 1 is a graph showing results from a competitive HO-1 ELISA onplasma from subjects, by group. AD group had significantly decreasedheme oxygenase-1 (HO-1) levels. No controls had a level below 1.0 ìg/ml.Mean values are indicated by the horizontal bars and statisticallysignificant differences between groups are indicated on the graph.

[0012]FIG. 2 is a graph showing results from a competitive HO-1 ELISA onlymphocytes from subjects, by group. AD group had significantlydecreased heme oxygenase-1 (HO-1) levels. No controls had a level below1.0 ìg/ml. Mean values are indicated by the horizontal bars andstatistically significant differences between groups are indicated onthe graph.

[0013]FIG. 3 is a graph showing results from a competitive HO-1 ELISA onplasma from subjects, by group, measured as % maximum inhibition, whichis directly proportional to protein concentrations. AD group hadsignificantly decreased % inhibition and therefore of heme oxygenase-1(HO-1) levels. Mean values are indicated by the horizontal bars andstatistically significant differences between groups are indicated onthe graph.

[0014]FIG. 4 is a graph showing HO-1 levels as a function of FolsteinMMSE scores in AD. Those with more severe dementia (lower MMSE) wereassociated with decreased HO-1 levels.

[0015]FIG. 5 is a graph showing HO-1 levels as a function of rate ofdecline in AD. The rate of global deterioration (x-axis) was derivedusing the difference between two successive Folstein mini-mental scores,multiplied by 3, divided by the intervening months. Those with morerapid decline (x>1) were associated with the most decreased HO-1 levels.

[0016]FIG. 6 is a Northern analysis of HO-1 mRNA levels in lymphocytesderived from subjects with Alzheimer's disease (AD), age-associatedcognitive decline (AACD) and normal elderly controls (NEC), etc. asdescribed in Example 3. Control GAPDH bands used to ensure uniformity ofRNA loading are depicted below the HO-1 bands.

[0017]FIG. 7 is a graph showing results from a Northern analysis of HO-1mRNA levels in lymphocytes derived from the subjects described inExample 3. AD group had significantly decreased heme oxygenase-1 (HO-1)mRNA levels.

DESCRIPTION OF THE EMBODIMENTS OF THE INVENTION

[0018] The Applicant has devised a diagnostic method, potentially usefulin the prediction, diagnosis, and prognostication of AD, AACD, andrelated neurological diseases. This diagnostic method is based on thediscovery that patients suffering from AD have a significantly lowerconcentration of heme oxygenase-1 (HO-1) in their lymphocytes andplasma, and, accordingly, a significantly lower concentration ofnucleotide sequences encoding HO-1 in their lymphocytes.

[0019] HO-1

[0020] Heme oxygenase-1 (HO-1) is an enzyme that catalyses the rapiddegradation of heme to biliverdin in brain and other tissues. This 32kDa member of the heat shock protein superfamily contains a heat-shockelement in its promoter region and is rapidly up regulated in responseto oxidative stress, metal ions, amino acid analogues, sulfhydrylagents, and hyperthermia. In response to oxidative stress, induction ofHO-1 may result in protection of cells by catabolizing pro-oxidantmetalloporphyrins, such as heme, into bile pigments (biliverdin,bilirubin), with free radical scavenging capabilities. Heme and otherintracellular ferrous iron chelates are capable of converting hydrogenperoxide to the highly cytotoxic hydroxyl radical.

[0021] Using immunostaining techniques in conjunction with laserscanning confocal microscopy, intense HO-1 immunoreactivity in neuronsand astrocytes of post-mortem hippocampus and temporal cortex derivedfrom AD subjects has been observed, whereas neural HO-1 staining wasfaint or non-existent in the hippocampus and temporal cortex of controlspecimens matched for age and post-mortem interval (Schipper et al.,1995). Furthermore, consistent co-localization of HO-1 toneurofibrillary tangles and senile plaques in the AD specimens has beendemonstrated. Finally, robust 32 kDa bands corresponding to HO-1 wereobserved by Western blotting of protein extracts derived from ADtemporal cortex and hippocampus after SDS-PAGE, whereas control HO-1bands were faint or absent. These results indicate that HO-1 issignificantly over-expressed in neurons and astrocytes of AD hippocampusand cerebral cortex relative to control brains and support thecontention that AD-affected tissues are experiencing chronic oxidativestress.

[0022] AACD

[0023] AACD is a term used to identify individuals who experience acognitive decline that falls short of dementia. Satisfaction of criteria(World Health Organization) for this diagnosis requires a report by theindividual or family of a decline in cognitive function, which isgradual, and present at least 6 months. There may be difficulties acrossany cognitive domains (although memory is impaired in the vast majorityof cases), and these must be supported by abnormal performance onquantitative cognitive assessments for which age and education norms areavailable for relatively healthy individuals (ie., the patient iscompared to normal subjects his/her own age). Performance must be atleast 1 SD below the mean value for the appropriate population on suchtests. Neither dementia, nor significant depression or drug effects maybe present. No cerebral or systemic disease or condition known to causecerebral cognitive dysfunction may be present. In the Applicant'sexperience, all patients who were classified as CDR.5 (“questionabledementia”) on the Clinical Dementia rating Scale and who met theseexclusions, also met the criteria for AACD.

[0024] About ⅓ of Alzheimer's patients have had a clearly definableperiod of isolated memory deficit which preceded their more globalcognitive decline (Haxby et al., 1992). Using AACD criteria which lookat other domains in addition to memory, the percentage with anidentifiable prodrome is likely higher. Fortunately, not all AACDindividuals seem to decline. It appears that a significant number ofthese subjects show a stable, non-progressive memory deficit on testing.

[0025] Related Disorders

[0026] Determining HO-1 concentration can also assist in predicting,diagnosing, or prognosticating other dementing diseases having similarmanifestations and/or in distinguishing such diseases from AD. Suchother diseases include Progressive Supranuclear Palsy, Vascular (i.e.multi-infarct) Dementia, Lewy Body Dementia, Huntington's Disease,Down's syndrome, normal pressure hydrocephalus, corticobasal ganglionicdegeneration, multisystem atrophy, head trauma, Creutzfeld-Jacobdisease, viral encephalitis and hypothyroidism.

EXAMPLE 1

[0027] Determination of Blood HO-1 Levels

[0028] Whole blood is collected from subjects in heparinized tubes. Thisis then layered over a Ficol Paque density gradient and centrifuged at1800 rpm for 20 minutes. The top plasma layer is then collected andsaved for ELISA after addition of Complete™ protease inhibitor cocktail(Boehringer Mannheim).

[0029] Lymphocyte Crude Microsomal Fraction

[0030] The layer of Ficol Paque gradient containing lymphocytes iscollected and washed three times in PBS and centrifuged at 3500 rpm for10 minutes. The resulting pellet is stored in Cell Culture FreezingMedium-DMSO (Gibco). After thawing, the pellet is washed three times inPBS and spun at 3000 rpm for 10 minutes. The resulting pellet isresuspended in 100 ìl 5 mM EDTA, 10 mM Tris and 150 mM NaCl pH 7.4 tolyse the cells. Lysate is homogenized and centrifuged at 100,000 rpm for50 minutes to pack down membrane fraction. Pellet is then solubilizedwith Triton-X100 and Tris pH 7.4.

[0031] HO-1 protein and HO-1 antibody (StressGen, Victoria BC) are usedto measure the percent competitive inhibition of HO-1 protein in humanplasma and lymphocytes. Alkaline phosphatase conjugated IgG (Bio Can)with p-nitrophenyl phosphate disodium (Sigma) as substrate is used tovisualize the reaction.

[0032] ELISA

[0033] Competitive ELISA for the detection and quantification of HO-1 isperformed according to the procedure of Wang et al. (1991) with thefollowing modifications: Immulon II ELISA plates (Dynatech) are coated(50 ìl/well) with 0.5 ìg/ml HO-1 protein (dissolved in coating bufferingcontaining Na2CO3 and NaHCO3 pH 9.6) and incubated overnight at 4° C.Excess protein is then washed off with washing buffer containing Tris,NaCl and Tween-20. Wells are blocked with 3% bovine serum albumen (BSA)in coating buffer and incubated 2 hrs at 37° C. 75 ìl of plasma orsolubilized membrane is then mixed with 75 ìl of HO-1 antibody (diluted1/50,000) and incubated for 2 hrs. at room temperature. After blocking,wells are washed with 50 ìl/well of plasma or solubilized membrane andHO-1 antibody mixture is added and incubated overnight at 4° C. Anyavailable antibody combines with the antigen exposed on the coated well.Secondary antibody consisting of alkaline phosphatase conjugated IgGanti-rabbit (diluted 1/1000) in 0.1% BSA is added and incubated 2 hrs at37° C. 50 ìl of the substrate (p-nitrophenyl phosphate disodiumdissolved in diethanolamine pH 9.8) is added to each well and incubatedfor 20 minutes at room temperature. The resulting colour reaction isread at 405 nm wavelength on a multi-well scanning spectrophotometer(Molecular Devices Corp.). The standard curve consists of equal volumesof HO-1 protein (10, 5.0, 2.5, 1.0, 0.5, 0.1 ìg/ml) and HO-1 antibody(1/50,000) which are mixed and incubated for 2 hrs at room temperature.This mixture is then added to the protein coated, BSA blocked wells inthe same manner as test samples. The results of % maximum inhibition arecalculated based on the standard curve generated by the known amounts ofHO-1 proteins in the standard samples.

EXAMPLE 2

[0034] Comparison of Blood HO-1 Levels Among Individuals With VaryingDegrees of Dementia

[0035] HO-1 levels in plasma and lymphocytes were assessed in normalelderly controls (NEC), patients probably with Alzheimer's Disease (AD)diagnosed according to NINCDS criteria, individuals with age-associatedcognitive decline not meeting AD criteria (AACD), and subjects withother neurological and medical disorders (Schipper et al., 1997;Schipper et al., in press). Demographic data is summarized in Table 1.

[0036] A competitive ELISA was used to determine HO-1 levels in plasmaand blood lymphocytes. In an early study (Schipper et al., 1997),Applicant found that mean plasma and lymphocyte concentrations of HO-1were significantly lower in AD subjects than in normal controls. Forplasma, the AD subjects had mean 0.715+/0.113 ìg/ml, vs 1.8+/−0.41 ìg/mlfor controls. This difference was highly significant (p=0.0012). Forlymphocytes, the AD subjects had HO-1 levels of 1.16+/−0.15 ìg/ml, vs2.23+/−0.56 ìg/ml for controls, again demonstrating a significantdifference (p<0.05). The AACD group overall demonstrated levels midwaybetween the two other groups: mean 1.01+/−0.35 ìg/ml for plasma, andmean 1.76+/−0.46 ìg/ml for lymphocytes. Subsequent studies (Schipper etal., in press) confirmed these results. See FIGS. 1, 2 and 3.

[0037] While there does exist overlap in scores between the groups,cut-off levels can be established (eg., 1.0 ìg/ml) below which no normalsubjects exist, but below which lie 25% of AD subjects, and also 25% ofAACD subjects. Presumably, any AACD subject lying in this range wouldhave a high (or perhaps even a certain) chance of progressing to AD overlongitudinal follow-up. In addition, analysis of the clinical data onthese subjects revealed important correlations. There was no correlationbetween HO-1 levels and age or duration of symptoms. There was, however,a significant correlation between HO-1 serum levels, and the FolsteinMini-mental state level, a general measure of dementia severity (r=0.57,p<0.005) (see FIG. 4). This suggests that as the AD subjects became moreseverely demented, their HO-1 levels progressively decrease.

[0038] Further analysis, however, suggests an alternative explanation.It was noted in the study that the most severely demented patients alsoseemed to be those who had deteriorated most rapidly. A measure of rateof global deterioration was derived (the difference between twosuccessive Folstein Mini-mental scores, multiplied by 3, divided by theintervening months). This formula was derived in view of the Applicant'spast experience that the typical AD subject declines by 3 or 4 points onthe MMSE per year. The association between rate of decline and HO-1levels was the most robust of all (r=−0.72, p<0.0001) (see FIG. 5). Theimplication is that the patients showing the most rapid declinedemonstrated the lowest levels of HO-1. Accordingly, HO-1 serum levelsoffer the possibility of producing useful prognostic information; thepresence of a low HO-1 level would imply more rapid deterioration overtime in AD subjects. The method can, accordingly, usefully be used tocompare HO-1 levels in the same patient over time in order toprognosticate the disease.

EXAMPLE 3

[0039] Comparison of Lymphocyte HO-1 mRNA Levels Among Individuals WithVarying Degrees of Dementia

[0040] Lymphocyte HO-1 mRNA levels were determined by Northern analysisin normal elderly controls (NEC), patients probably with Alzheimer'sDisease (AD) diagnosed according to NINCDS criteria, individuals withage-associated cognitive decline not meeting AD criteria (AACD), andsubjects with other neurological and medical disorders (Schipper et al.,1997; Schipper et al., in press) derived from the same sources anddiagnosed according to the same criteria as described in Example 2above.

[0041] Lymphocyte fractions were obtained by differential centrifugationof whole blood on Ficol Paque gradients as described above. CytoplasmicRNA was isolated from the lymphocytes using an acid guanidiniumthiocyanate-phenol-chloroform extraction method (Chomezynski P et al.,1997). Six micrograms of RNA was denatured and size-separated byelectrophoresis on 1% agarose/formaldehyde gels. RNA integrity wasconfirmed by ethidium bromide staining. The RNA was transferred ontoHybond-N filter paper and covalently cross-linked to the membrane by UVlight for two minutes. The hybridization probe (HO-1; 1.0 kb) wasprepared by random priming using the Random Primer DNA Labelling System(Feinberg A P et al., 1984). Prehybridization was performed for 12 hoursat 42° C. in a buffer containing formamide deionized, 5×Denhardt'sreagent, 6×SSPE and 0.5% SDS. The hybridization buffer consisted of theprehybridization buffer without 5×Denhardt's reagent, and 32P-labelleddenatured DNA probe (Noonberg S B et al., 1994). Equal loading of RNAwas confirmed by hybridization with a cDNA for the (housekeeping) gene,glyceraldehyde-3-phosphate dehydrogenase (GAPDH). All washes wereperformed under stringent conditions (1×SSC and 0.2% SDS for 45 minutesat room temperature, 0.4×SSC and 0.2% SDS for 15 minutes at 65° C.). TheRNA hybridizing with cDNA probes was visualized by autoradiography usingan intensifying screen at −80° C. (Church G M et al., 1984).

[0042] Lymphocyte HO-1 mRNA bands, assayed by Northern analysis, wereuniformly robust in the NEC subjects. In contrast, HO-1 mRNA bands werefaint or undetectable in lymphocytes derived from AD subjects and ofvariable intensity in the AACD cohort (see FIG. 6). Quantification bydensitometry produced the results in FIG. 7.

[0043] The above examples illustrate the testing method using blood andlymphocytes as the tissue samples. The method can also be applied toother tissues such as cerebrospinal fluid and to fibroblast cell linesderived from patients.

[0044] The examples also illustrate application of the testing method todetect concentration of HO-1 as well as of HO-1 mRNA.

[0045] The test can be applied to compare the concentration of HO-1 or anucleotide sequence encoding it in a specific patient over a period oftime, or to compare the concentration of HO-1 or a nucleotide sequenceencoding it in a patient with the corresponding concentration in anormal control population.

[0046] A kit or commercial package for carrying out the testing methodincludes a means for determining the concentration of heme oxygenase-1(HO-1) and/or a nucleotide sequence encoding HO-1, in tissue obtainedfrom a patient. It also includes instructions for comparing saidconcentration with an established standard of the correspondingconcentration of HO-1 and/or an HO-1 encoding nucleotide sequence incorresponding tissue obtained from at least one normal elderly controlperson or from the patient at another time. TABLE 1 AGE SEX MMSE EDUC.DIAGNOSIS II (mean) F/M (mean) (mean) CONTROL 31 62.5 17/14 29.4 14.1Age 40-60(NYC)⊕ 7 47.1 1/6 30 16.6 60 +(NBC) * 24 77.9 16/8  28.9 11.7AACD * 25 76.3 15/10 26.9 10.5 AD * 50 76.8 27/23 21.1 10.9 NAD *♦ 1674.3 6/9 11.5 LBD 4 71.25 2/2 18.2 — CBGD 2 69.6 1/1 7.5 — HD 1 63 M 24NPH 2 81.0 2M 25 — MID 4 77.5 1/3 6.2 — MSA 1 86 M 11 — DOWN 1 55 F 0 —HT 1 86 F 0 — PD  21 68.7 10/11 29.0 — ALS Δ 14 55.7 8/6 28.9 — SALS 1356.1 8/5 28.8 — FALS 1 50 M 30 — STR # 5 61.8 1/4 26 — RD

8 63.4 6/2 27.6 — LIV

5 51.8 2/3 27.2 — HS 4 51.0 2/2 26.5 — CAH 1 55 M 30 —

REFERENCES

[0047] Chomczynski P and Sacchi N. (1987) Single-step method of RNAisolation by acid guanidinium thiocyanate-phenol-chlorophorm extraction.Analytical Biochem 162:156-159.

[0048] Church G M and Gilbert W. (1984) Genomic sequencing. Proc Nat AcaSci USA 81:1991-1995.

[0049] Feinberg A P and Vogelstein B. (1984) A Technique forradiolabelling DNA restriction endonuclease fragments to high specificactivity. Analytical Biochem 137:266-267.

[0050] Grossi, D., et al. (1988) Senile dementias. II InternationalSymposium (pp. 97-99). Paris: John Libbey Eurotext.

[0051] Haxby J V, et al. (1992) Individual trajectories of cognitivedecline in patients with dementia of the Alzheimer type. J Clin ExpNeuropsychol 14:575-592.

[0052] Mann, U., et al. (1992) Heterogeneity in Alzheimer's disease:Progression rate segregated by distinct neuropsychological and cerebralmetabolic profiles. J Neurol Neurosurg Psychiatry 55:956-959.

[0053] Noonberg S B, et al. (1994) Detection of triplex-forming RNAoligonucleotides by triplex blotting. BioTechniques 16:1070-1072.

[0054] Schipper H M, et al. (1995). Expression of heme oxygenase-1 inthe senescent and Alzheimer-diseased brain. Ann Neurol 37:758-768.

[0055] Schipper H M, et al. (1997) Blood Heme Oxygenase Levels aredecreased in Alzheimer's disease. Proc Soc for Neuroscience 123:1-41.

[0056] Schipper H M, et al. (in press) Blood Heme Oxygenase-1 Levels areDecreased in Alzheimer's Disease and Correlate Inversely with Rates ofDisease Progression.

[0057] Schipper H M, et al. (in press) Further Evaluation of HemeOxygenase-1 as a Biological Marker in Alzheimer's Disease.

[0058] Wang, G. P., et al. (1991) Alzheimer's disease: paired helicalfilament immunoreactivity in cerebrospinal fluid. Acta Neuropathol82:6-12.

1. A commercial package comprising; means for determining theconcentration of heme oxygenase-1 (HO-1) and/or a nucleotide sequenceencoding HO-1, in bodily fluid or non-neural tissue obtained from apatient; and instructions for assessing a dementing disease in thepatient; wherein said commercial package is used to predict the onsetof, diagnose, or prognosticate a dementing disease.
 2. A commercialpackage comprising; means for determining the concentration of hemeoxygenase-1 (HO-1) and/or a nucleotide sequence encoding HO-1, in bodilyfluid or non-neural tissue obtained from a patient; and instructions forcomparing said concentration with the corresponding concentration ofHO-1 and/or an HO-1 encoding nucleotide sequence in corresponding bodilyfluid or non-neural tissue obtained from at least one control person;wherein a reduced concentration is used to predict the onset of,diagnose, or prognosticate an Alzheimer dementing disease; and wherein aconcentration that is not reduced indicates that the dementing diseaseis not an Alzheimer dementing disease.
 3. The commercial packageaccording to claim 1 wherein the means is for determining theconcentration of HO-1 and the bodily fluid is selected from plasma andcerebrospinal fluid and the tissue is selected from lymphocytes andfibroblasts, or the means is for determining the concentration of HO-1encoding nucleotide sequence and the tissue is selected from lymphocytesand fibroblasts.
 4. The commercial package according to claim 2 whereinthe means is for determining the concentration of HO-1 and the bodilyfluid is selected from plasma and cerebrospinal fluid and the tissue isselected from lymphocytes and fibroblasts, or the means is fordetermining the concentration of HO-1 encoding nucleotide sequence andthe tissue is selected from lymphocytes and fibroblasts.
 5. A commercialpackage according to claim 3 wherein the bodily fluid is plasma.
 6. Acommercial package according to claim 4 wherein the bodily fluid isplasma.
 7. The commercial package according to claim 3 wherein the meansis for determining the concentration of HO-1 and the tissue islymphocytes.
 8. The commercial package according to claim 4 wherein themeans is for determining the concentration of HO-1 and the tissue islymphocytes.
 9. The commercial package according to claim 3 wherein themeans is for determining the concentration of HO-1 mRNA and the tissueis lymphocytes.
 10. The commercial package according to claim 4 whereinthe means is for determining the concentration of HO-1 mRNA and thetissue is lymphocytes.
 11. The commercial package according to claim 1wherein the dementing disease is selected from the group consisting ofAlzheimer's Disease, Age-Associated Cognitive Decline, ProgressiveSupranuclear Palsy, Vascular (i.e. multi-infarct) Dementia, Lewy BodyDementia, Huntington's Disease, Down's syndrome, normal pressurehydrocephalus, corticobasal ganglionic degeneration, multisystematrophy, head trauma, Creutzfeld-Jacob disease, viral encephalitis andhypothyroidism.
 12. The commercial package according to claim 2 whereinthe dementing disease is selected from the group consisting ofAlzheimer's Disease, Age-Associated Cognitive Decline, ProgressiveSupranuclear Palsy, Vascular (i.e. multi-infarct) Dementia, Lewy BodyDementia, Huntington's Disease, Down's syndrome, normal pressurehydrocephalus, corticobasal ganglionic degeneration, multisystematrophy, head trauma, Creutzfeld-Jacob disease, viral encephalitis andhypothyroidism.
 13. The commercial package according to claim 2 whereinthe at least one control person is a normal age-matched person.
 14. Thecommercial package according to claim 2 wherein the at least one controlperson is the patient from whom the corresponding concentration of HO-1and/or an HO-1 encoding nucleotide sequence in bodily fluid andnon-neural tissue was obtained at an earlier date, and the commercialpackage is used to prognosticate a dementing disease.
 15. A commercialpackage comprising means for determining the concentration of hemeoxygenase-1 (HO-1) and/or a nucleotide sequence encoding HO-1, in bodilyfluid or tissue obtained from a patient, and instructions for comparingsaid concentration with an established standard of the correspondingconcentration of HO-1 and/or an HO-1 encoding nucleotide sequence incorresponding bodily fluid or tissue obtained from at least one normalage-matched control person or from the patient at an earlier time. 16.The commercial package according to claim 15 wherein the means is fordetermining the concentration of HO-1 and the bodily fluid is selectedfrom plasma and cerebrospinal fluid and the tissue is selected fromlymphocytes and fibroblasts, or the means is for determining theconcentration of HO-1 encoding nucleotide sequence and the tissue isselected from lymphocytes and fibroblasts.
 17. A commercial packageaccording to claim 16 wherein the bodily fluid is plasma.
 18. Thecommercial package according to claim 16 wherein the means is fordetermining the concentration of HO-1 or HO-1 mRNA and the tissue islymphocytes.
 19. The commercial package of claim 15 wherein thecorresponding bodily fluid or tissue is obtained from at least onenormal age-matched control person.
 20. The commercial package of claim15 wherein the corresponding bodily fluid or tissue is obtained from thepatient at an earlier time.